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Bergamo 2 two photon microscope

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The Bergamo II is a two-photon microscope designed for high-resolution imaging of biological samples. It utilizes the principle of two-photon excitation to enable deep tissue penetration and reduced photodamage compared to single-photon microscopy. The core function of the Bergamo II is to provide researchers with a versatile and powerful tool for advanced fluorescence imaging applications.

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Bergamo II (18 citations) Bergamo II microscope (17 citations) Bergamo two-photon microscope (2 citations) Bergamo 2-photon microscope (2 citations) Two-photon microscope (2 citations)

4 protocols using bergamo 2 two photon microscope

1

In Vivo Two-Photon Imaging of Neuronal Activity

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Experimental animals were placed head-fixed on a wheel under light isoflurane sedation, then allowed to wake up. The mouse was acclimated to the wheel in a free running mode for 10 min, head-fixed under a Thorlabs Bergamo II two-photon microscope (Thorlabs, NJ) with a Nikon 25X (NA 1.1) water-immersion objective lens (Nikon Instruments Inc., NY). Time-lapse images were acquired with Thorlabs software at 6–7 frames/sec for 4630 frames for 12 min (2 min for each speed for both increasing and decreasing mode). The interval between two modes at each trial is 1 min. The laser power was adjusted to the minimum necessary to achieve ideal fluorescence intensities during each imaging session. On average, about 20–30 mW of power arrives at the sample. The scanning depth is ~ 200 µm from the dura. After completing the experiment, the animals were perfused with 10% formaldehyde, and the brain was sectioned to confirm the imaging depth and the location of the viral expression.
Yue Y., Ash R.T., Boyle N., Kinter A., Li Y., Zeng C, & Lu H. (2022). MeCP2 deficiency impairs motor cortical circuit flexibility associated with motor learning. Molecular Brain, 15, 76.
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2

Two-Photon Microscopy Imaging Protocol

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Imaging was performed using either a galvanometric-scanner based MOM (Sutter) or a resonant-scanner based (8 kHz) Bergamo II two-photon microscope (Thorlabs), both controlled by ScanImage (Vidrio). Using the MOM system, we acquired images of 128 × 128 pixels at a single depth at 5.92 Hz frame rate. With the Bergamo II, we acquired images of 380 × 512 pixels at 1 or 4 depths at 40 Hz or 8 Hz frame rate, respectively. We obtained similar results with both systems, so all data were pooled. The illumination light source was a Ti:sapphire laser (Chameleon Ultra II, Coherent) used at an excitation wavelength of 910 nm. The laser power under the objective (16×, Nikon) was typically set to 30 mW and never exceeded 50 mW (laser pulse width 140 fs at a repetition rate of 80 MHz).
Keller A.J., Dipoppa M., Roth M.M., Caudill M.S., Ingrosso A., Miller K.D, & Scanziani M. (2020). A Disinhibitory Circuit for Contextual Modulation in Primary Visual Cortex. Neuron, 108(6), 1181-1193.e8.
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3

Two-Photon Microscopy for Cell Imaging

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Imaging was performed using either a galvanometric-scanner based MOM (Sutter) or a resonant-scanner based (8 kHz) Bergamo II two-photon microscope (Thorlabs), both controlled by ScanImage (Vidrio). Using the MOM system, we acquired images of 128 × 128 pixels at a single depth at 5.92 Hz frame rate. With the Bergamo II, we acquired images of 380 × 512 pixels at 1 or 4 depths at 40 Hz or 8 Hz frame rate, respectively. We obtained similar results with both systems, so all data were pooled. The illumination light source was a Ti:sapphire laser (Chameleon Ultra II, Coherent) used at an excitation wavelength of 910 nm for green indicator imaging and of 1040 nm for red indicator imaging. The laser power under the objective (16×, Nikon) never exceeded 50 mW (laser pulse width 140 fs at a repetition rate of 80 MHz).
Keller A.J., Roth M.M, & Scanziani M. (2020). Feedback Generates a Second Receptive Field in Neurons of Visual Cortex. Nature, 582(7813), 545-549.
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4

Two-Photon Microscopy for Cell Imaging

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Imaging was performed using either a galvanometric-scanner based MOM (Sutter) or a resonant-scanner based (8 kHz) Bergamo II two-photon microscope (Thorlabs), both controlled by ScanImage (Vidrio). Using the MOM system, we acquired images of 128 × 128 pixels at a single depth at 5.92 Hz frame rate. With the Bergamo II, we acquired images of 380 × 512 pixels at 1 or 4 depths at 40 Hz or 8 Hz frame rate, respectively. We obtained similar results with both systems, so all data were pooled. The illumination light source was a Ti:sapphire laser (Chameleon Ultra II, Coherent) used at an excitation wavelength of 910 nm for green indicator imaging and of 1040 nm for red indicator imaging. The laser power under the objective (16×, Nikon) never exceeded 50 mW (laser pulse width 140 fs at a repetition rate of 80 MHz).
Keller A.J., Roth M.M, & Scanziani M. (2020). Feedback Generates a Second Receptive Field in Neurons of Visual Cortex. Nature, 582(7813), 545-549.
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